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Title: Leucine-rich-repeat-containing variable lymphocyte receptors as modules to target plant-expressed proteins

Abstract

The ability to target and manipulate protein-based cellular processes would accelerate plant research; yet, the technology to specifically and selectively target plant-expressed proteins is still in its infancy. Leucine-rich repeats (LRRs) are ubiquitously present protein domains involved in mediating protein–protein interactions. LRRs confer the binding specificity to the highly diverse variable lymphocyte receptor (VLR) antibodies (including VLRA, VLRB and VLRC types) that jawless vertebrates make as the functional equivalents of jawed vertebrate immunoglobulin-based antibodies. Here, VLRBs targeting an effector protein from a plant pathogen, HopM1, were developed by immunizing lampreys and using yeast surface display to select for high-affinity VLRBs. HopM1-specific VLRBs (VLRM1) were expressed in planta in the cytosol, the trans-Golgi network, and the apoplast. Expression of VLRM1 was higher when the protein localized to an oxidizing environment that would favor disulfide bridge formation (when VLRM1 was not localized to the cytoplasm), as disulfide bonds are necessary for proper VLR folding. VLRM1 specifically interacted in planta with HopM1 but not with an unrelated bacterial effector protein while HopM1 failed to interact with a non-specific VLRB. Later, VLRs may be used as flexible modules to bind proteins or carbohydrates of interest in planta, with broad possibilities for their use bymore » binding directly to their targets and inhibiting their action, or by creating chimeric proteins with new specificities in which endogenous LRR domains are replaced by those present in VLRs.« less

Authors:
 [1];  [1];  [2];  [2];  [3]
  1. Michigan State Univ., East Lansing, MI (United States). DOE Plant Research Lab.
  2. Emory Univ., Atlanta, GA (United States). Dept. of Pathology and Lab. Medicine
  3. Michigan State Univ., East Lansing, MI (United States). DOE Plant Research Lab., Dept. of Plant Biology, Plant Resilience Inst., and Howard Hughes Medical Inst.
Publication Date:
Research Org.:
Michigan State Univ., East Lansing, MI (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
OSTI Identifier:
1368384
Grant/Contract Number:  
FG02-91ER20021
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Plant Methods
Additional Journal Information:
Journal Volume: 13; Journal Issue: 1; Journal ID: ISSN 1746-4811
Publisher:
BioMed Central
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; Protein targeting; Leucine-rich repeat; Variable lymphocyte receptor; Modules HopM1

Citation Formats

Velásquez, André C., Nomura, Kinya, Cooper, Max D., Herrin, Brantley R., and He, Sheng Yang. Leucine-rich-repeat-containing variable lymphocyte receptors as modules to target plant-expressed proteins. United States: N. p., 2017. Web. doi:10.1186/s13007-017-0180-8.
Velásquez, André C., Nomura, Kinya, Cooper, Max D., Herrin, Brantley R., & He, Sheng Yang. Leucine-rich-repeat-containing variable lymphocyte receptors as modules to target plant-expressed proteins. United States. doi:10.1186/s13007-017-0180-8.
Velásquez, André C., Nomura, Kinya, Cooper, Max D., Herrin, Brantley R., and He, Sheng Yang. Wed . "Leucine-rich-repeat-containing variable lymphocyte receptors as modules to target plant-expressed proteins". United States. doi:10.1186/s13007-017-0180-8. https://www.osti.gov/servlets/purl/1368384.
@article{osti_1368384,
title = {Leucine-rich-repeat-containing variable lymphocyte receptors as modules to target plant-expressed proteins},
author = {Velásquez, André C. and Nomura, Kinya and Cooper, Max D. and Herrin, Brantley R. and He, Sheng Yang},
abstractNote = {The ability to target and manipulate protein-based cellular processes would accelerate plant research; yet, the technology to specifically and selectively target plant-expressed proteins is still in its infancy. Leucine-rich repeats (LRRs) are ubiquitously present protein domains involved in mediating protein–protein interactions. LRRs confer the binding specificity to the highly diverse variable lymphocyte receptor (VLR) antibodies (including VLRA, VLRB and VLRC types) that jawless vertebrates make as the functional equivalents of jawed vertebrate immunoglobulin-based antibodies. Here, VLRBs targeting an effector protein from a plant pathogen, HopM1, were developed by immunizing lampreys and using yeast surface display to select for high-affinity VLRBs. HopM1-specific VLRBs (VLRM1) were expressed in planta in the cytosol, the trans-Golgi network, and the apoplast. Expression of VLRM1 was higher when the protein localized to an oxidizing environment that would favor disulfide bridge formation (when VLRM1 was not localized to the cytoplasm), as disulfide bonds are necessary for proper VLR folding. VLRM1 specifically interacted in planta with HopM1 but not with an unrelated bacterial effector protein while HopM1 failed to interact with a non-specific VLRB. Later, VLRs may be used as flexible modules to bind proteins or carbohydrates of interest in planta, with broad possibilities for their use by binding directly to their targets and inhibiting their action, or by creating chimeric proteins with new specificities in which endogenous LRR domains are replaced by those present in VLRs.},
doi = {10.1186/s13007-017-0180-8},
journal = {Plant Methods},
number = 1,
volume = 13,
place = {United States},
year = {Wed Apr 19 00:00:00 EDT 2017},
month = {Wed Apr 19 00:00:00 EDT 2017}
}

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